上海农业学报2017，Vol.33Issue(2)：89-95,7.DOI:10.15955/j.issn1000-3924.2017.02.17
猪圆环病毒2型荧光定量PCR检测试剂盒的研制及应用
Development and application of fluorescent quantitation PCR detection kit of porcine circovirus type 2
摘要
Abstract
In order to develop the diagnostic technique and diagnostic kit of porcine circovirus type 2 (PCV2),specific primers and a probe for the PCV2 ORF2 gene were specially designed,and the PCV2 was detected by a fluorescent quantitation PCR method.The tests showed that the cycle threshold Ct and the standard DNA template had an excellent linear relationship between 101-107 copies per microliter,and their correlation coefficient was 0.9999;The variation coefficient of Ct values was 0.59％-1.05％ in the intra-lot test and 1.9％-4.2％ in the inter-lot test,there being a good repeatability;The developed detection kit had no cross reaction with hog cholera virus (HCV),porcine respiratory and reproductive syndrome virus (PRRSV)and swine pseudorabies virus (PRV),and its detection and quantitation limits were 10 and 100 copy numbers respectively.The detection of 1 000 clinical samples by this method showed that the 710 samples were positive.The result indicated that this kit was high in sensitiveness,strong in specificity and good in stability,providing a simple and rapid method for PCV2 monitoring and epidemiological survey.关键词
猪圆环病毒2型/荧光定量/PCR检测/诊断试剂盒Key words
Porcine circovirus type 2/Fluorescent quantitation/PCR detection/Diagnostic kit分类
农业科技引用本文复制引用
任方方,胡瑞丽,倪建平,许燕,李春华,赵笑,杜亚楠,赵凯..猪圆环病毒2型荧光定量PCR检测试剂盒的研制及应用[J].上海农业学报,2017,33(2):89-95,7.基金项目
上海市闵行区重大产业化专项(2014MH076) （2014MH076）
